Dual function engineered t cells with hpv e6 specificity and pd-1 blockade

Abstract

The present invention generally relates to engineered cells and compositions thereof, particularly, T cells comprising genetically engineered T Cell receptors (TCRs) and checkpoint inhibitors (CPIs). Methods for using the compositions to treat cancer are also disclosed herein. Genetically engineered T cells that recognize tumor antigen HPV E6 and simultaneously secrete a single-chain antibody that blocks Programmed Cell Death Protein 1 (PD-1). Also provided is an immunotherapy for HPV E6 expression related cancers.

1 . An engineered T cell, comprising:
 a nucleic acid encoding (a) genetically engineered antigen receptor that specifically binds to an antigen from HPV; and   (b) an inhibitory protein that reduces function or expression of inhibitory receptors in a tumor.   
     
     
         2 . The engineered T cell of  claim 1 , wherein the antigen comprises E6 or E7. 
     
     
         3 . The engineered T cell of  claim 1 , wherein tumor target comprises one or more of PD-1. 
     
     
         4 . The engineered T cell of  claim 3 , wherein the inhibitory protein is an anti PD1 antibody. 
     
     
         5 . The engineered T cell of  claim 4 , wherein the anti PD1 antibody is a single chain antibody. 
     
     
         6 . The engineered T cell of  claim 5 , wherein the anti PD1 antibody comprised motif sequences 1) heavy chain CDR1 of GYTFTNYY, CDR2 of INPSNGGT, and CDR3 of TRRDYNYDGGFDY; 2) Light chain CDR1 of KSVSTSGFN, CDR2 of LAS and CDR3 of QHGRELPLT. 
     
     
         7 . The engineered T cell of any of  claims 1 - 6 , wherein the inhibitory nucleic acid molecule comprises a sequence complementary to a PD1-encoding nucleic acid. 
     
     
         8 . The engineered T cell of any of  claims 1 - 6 , wherein the inhibitory nucleic acid molecule comprises an antisense oligonucleotide complementary to a PD1-encoding nucleic acid. 
     
     
         9 . The engineered T cell of any of  claims 1 - 6 , wherein the inhibitory protein or antibody is constitutively expressed. 
     
     
         10 . The engineered T cell of  claim 9 , wherein the inhibitory protein is an antibody PD1 which is constitutively expressed. 
     
     
         11 . A nucleic acid comprising (a) a nucleic acid encoding genetically engineered antigen receptor that specifically binds to an antigen from HPV; and (b) an inhibitory nucleic acid molecule that reduces the expression of an inhibitory receptor in a tumor. 
     
     
         12 . The nucleic acid of  claim 11 , wherein the antigen receptor is E6 of HPV. 
     
     
         13 . The nucleic acid of  claim 12 , wherein the tumor target is PD1. 
     
     
         14 . Polypeptides encoded by the nucleic acid of any of  claims 11 - 13 . 
     
     
         15 . A vector comprising the nucleic acid of any of  claims 11 - 13 . 
     
     
         16 . The vector of  claim 15 , wherein vector is a retroviral vector. 
     
     
         17 . A method of producing a genetically engineered T cell, comprising introducing a vector comprising 1) a nucleic acid encoding genetically engineered antigen receptor that specifically binds to a first antigen into a population of cells comprising T cells, the first antigen receptor specifically target to E6 receptor of HPV, (b) a nucleic acid molecule encoding an inhibitory protein capable of leading to a reduction of expression of PD-1 or PD-L1 and/or inhibiting upregulation of PD-1 or PD-L1 in T cells in the population upon incubation under one or more conditions. 
     
     
         18 . A pharmaceutical composition, comprising the engineered T cell of any of  claims 1 - 10  and a pharmaceutically acceptable carrier. 
     
     
         19 . A method for treating cancer comprising administering to a subject in need thereof, a therapeutically effective amount of the pharmaceutical composition of  claim 18 . 
     
     
         20 . The method of  claim 19 , wherein the cancer is cervical cancer or head and neck cancer. 
     
     
         21 . The method of  claim 20 , further comprising administering to the subject a therapeutically effective amount of an existing therapy comprising chemotherapy or radiation. 
     
     
         22 . The method of  claim 21 , wherein the cell and the existing therapy are administered sequentially or simultaneously. 
     
     
         23 . The engineered T cell in  claim 1 , wherein the tumor comprises lymphocytes or tumor-infiltrating lymphocytes. 
     
     
         24 . The nucleic acid of  claim 11 , wherein the tumor comprises lymphocytes or tumor-infiltrating lymphocytes.